Crystals of SctV from different species reveal variable symmetry for the cytosolic domain of the type III secretion system export gate.

Type III secretion systems (T3SSs) are proteinaceous devices employed by Gram-negative bacteria to directly transport proteins into a host cell. Substrate recognition and secretion are strictly regulated by the export apparatus of the so-called injectisome. The export gate SctV engages chaperone-bound substrates of the T3SS in its nonameric cytoplasmic domain. Here, the purification and crystallization of the cytoplasmic domains of SctV from Photorhabdus luminescens (LscVC) and Aeromonas hydrophila (AscVC) are reported. Self-rotation functions revealed that LscVC forms oligomers with either eightfold or ninefold symmetry in two different crystal forms. Similarly, AscVC was found to exhibit tenfold rotational symmetry. These are the first instances of SctV proteins forming non-nonameric oligomers.

Early Stroke Induces Long-Term Impairment of Adult Neurogenesis Accompanied by Hippocampal-Mediated Cognitive Decline.

Stroke increases neurogenesis in the adult dentate gyrus in the short term, however, long-term effects at the cellular and functional level are poorly understood. Here we evaluated the impact of an early stroke lesion on neurogenesis and cognitive function of the aging brain. We hypothesized that a stroke disturbs dentate neurogenesis during aging correlate with impaired flexible learning. To address this issue a stroke was induced in 3-month-old C57Bl/6 mice by a middle cerebral artery occlusion (MCAO). To verify long-term changes of adult neurogenesis the thymidine analogue BrdU (5-Bromo-2'-deoxyuridine) was administrated at different time points during aging. One and half months after BrdU injections learning and memory performance were assessed with a modified version of the Morris water maze (MWM) that includes the re-learning paradigm, as well as hippocampus-dependent and -independent search strategies. After MWM performance mice were transcardially perfused. To further evaluate in detail the stroke-mediated changes on stem- and progenitor cells as well as endogenous proliferation nestin-green-fluorescent protein (GFP) mice were used. Adult nestin-GFP mice received a retroviral vector injection in the hippocampus to evaluate changes in the neuronal morphology. At an age of 20 month the nestin-GFP mice were transcardially perfused after MWM performance and BrdU application 1.5 months later. The early stroke lesion significantly decreased neurogenesis in 7.5- and 9-month-old animals and also endogenous proliferation in the latter group. Furthermore, immature doublecortin (DCX)-positive neurons were reduced in 20-month-old nestin-GFP mice after lesion. All MCAO groups showed an impaired performance in the MWM and mostly relied on hippocampal-independent search strategies. These findings indicate that an early ischemic insult leads to a dramatical decline of neurogenesis during aging that correlates with a premature development of hippocampal-dependent deficits. Our study supports the notion that an early stroke might lead to long-term cognitive deficits as observed in human patients after lesion.

Cortical spreading depolarization increases adult neurogenesis, and alters behavior and hippocampus-dependent memory in mice.

Cortical spreading depolarizations are an epiphenomenon of human brain pathologies and associated with extensive but transient changes in ion homeostasis, metabolism, and blood flow. Previously, we have shown that cortical spreading depolarization have long-lasting consequences on the brains transcriptome and structure. In particular, we found that cortical spreading depolarization stimulate hippocampal cell proliferation resulting in a sustained increase in adult neurogenesis. Since the hippocampus is responsible for explicit memory and adult-born dentate granule neurons contribute to this function, cortical spreading depolarization might influence hippocampus-dependent cognition. To address this question, we induced cortical spreading depolarization in C57Bl/6 J mice by epidural application of 1.5 mol/L KCl and evaluated neurogenesis and behavior at two, four, or six weeks thereafter. Congruent with our previous findings in rats, we found that cortical spreading depolarization increases numbers of newborn dentate granule neurons. Moreover, exploratory behavior and object location memory were consistently enhanced. Reference memory in the water maze was virtually unaffected, whereas memory formation in the Barnes maze was impaired with a delay of two weeks and facilitated after four weeks. These data show that cortical spreading depolarization produces lasting changes in psychomotor behavior and complex, delay- and task-dependent changes in spatial memory, and suggest that cortical spreading depolarization-like events affect the emotional and cognitive outcomes of associated brain pathologies.

Cyclin D2 knockout mice with depleted adult neurogenesis learn Barnes maze task.

There is a broad discussion concerning the function of new neurons in the adult brain. An increasingly accepted hypothesis proposes their crucial role in spatial learning. In this work, however, we demonstrate adult cyclin D2 knockout (cD2 KO) mice, which lack adult hippocampal neurogenesis, are able to learn a spatial version of the Barnes maze. Similar to wild type (WT) controls, these mutant mice exhibited several indicators of learning during 6 days of training: successively shorter latency and distance, higher speed, and decreasing number of errors. WT and cD2 KO mice showed improved search strategies, which became increasingly spatial. During probe Trial 1, mutant mice attained the highest significant number of nose-pokes at the former target hole compared with all the other holes. Both WT and cD2 KO mice covered shorter distances during probe Trial 2, whereas the mutant mice showed higher speed. We also discuss the possibility that some of the observed differences displayed by cD2 KO mice during training and at the probe trials-for example, longer mean distance and more errors-are associated with a smaller hippocampal formation. Our results suggest that adult brain neurogenesis is not obligatory for learning the Barnes maze.

Head capsule, chephalic central nervous system and head circulatory system of an aberrant orthopteran, Prosarthria teretrirostris (Caelifera, Hexapoda).

The head capsule, the circulatory system and the central nervous system of the head of Prosarthria teretrirostris (Proscopiidae) is described in detail, with special consideration of modifications resulting from the aberrant head shape. The transformations of the head are completely different from those found in phasmatodeans, which are also characterised by twig mimesis. The circulatory system is distinctly modified. A hitherto undescribed additional structure in the posterior head region very likely functions as a pulsatile organ. The cephalic central nervous system is strongly elongated, with changes in the position of the suboesophageal ganglion, the corpora cardiaca and the course of the nervus mandibularis. Three-dimensional reconstructions of these two organ systems in combination with the pharynx were made using Alias Maya 6.0 software. Comparisons with other representatives of Caelifera suggest a clade comprising Proscopiidae and Morabinae. The presence of a transverse muscle connecting the antennal ampullae in Prosarthria shows that this structure likely belongs to the groundplan of Orthoptera, even though it is missing in different representatives of this group. The transverse ampullary muscle is a potential synapomorphy of Orthoptera, Phasmatodea and Dictyoptera.